title: Differentiation of human induced pluripotent stem cells into functional enterocyte-like cells using a simple method.
abstract: Human induced pluripotent stem (iPS) cells were differentiated into the endoderm using activin A and were then treated with fibroblast growth factor 2 (FGF2) for differentiation into intestinal stem cell-like cells. These immature cells were then differentiated into enterocyte-like cells using epidermal growth factor (EGF) in 2% fetal bovine serum (FBS). At the early stage of differentiation, mRNA expression of caudal type homeobox 2 (CDX2), a major transcription factor related to intestinal development and differentiation, and leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5), an intestinal stem cell marker, was markedly increased by treatment with FGF2. When cells were cultured in medium containing EGF and a low concentration of FBS, mRNAs of specific markers of intestinal epithelial cells, including sucrase-isomaltase, the intestinal oligopeptide transporter SLC15A1/peptide transporter 1 (PEPT1), and the major metabolizing enzyme CYP3A4, were expressed. In addition, sucrase-isomaltase protein expression and uptake of β-Ala-Lys-N-7-amino-4-methylcoumarin-3-acetic acid (β-Ala-Lys-AMCA), a fluorescence-labeled substrate of the oligopeptide transporter, were detected. These results demonstrate a simple and direct method for differentiating human iPS cells into functional enterocyte-like cells.
authors.affiliation: Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Iwao
authors.forename: Masashi
authors.initials: M
authors.lastname: Toyota
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Miyagawa
authors.forename: Hajime
authors.initials: H
authors.lastname: Okita
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Kiyokawa
authors.forename: Hidenori
authors.initials: H
authors.lastname: Akutsu
authors.forename: Akihiro
authors.initials: A
authors.lastname: Umezawa
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Nagata
authors.forename: Tamihide
authors.initials: T
authors.lastname: Matsunaga
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002417:Cattle; D002454:Cell Differentiation; D002460:Cell Line; D003374:Coumarins / Q000378:metabolism; D051544:Cytochrome P-450 CYP3A / Q000235:genetics / Q000378:metabolism; D004151:Dipeptides / Q000378:metabolism; D004912:Erythrocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D009834:Oligo-1,6-Glucosidase / Q000235:genetics / Q000378:metabolism; D000074011:Peptide Transporter 1; D012333:RNA, Messenger / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D013393:Sucrase / Q000235:genetics / Q000378:metabolism; D027981:Symporters / Q000235:genetics / Q000378:metabolism
chemical_list: D015415:Biomarkers; D003374:Coumarins; D004151:Dipeptides; C116884:LGR5 protein, human; D000074011:Peptide Transporter 1; D012333:RNA, Messenger; D043562:Receptors, G-Protein-Coupled; C082186:SLC15A1 protein, human; D027981:Symporters; C434800:alanyl-lysyl-N(epsilon)-7-amino-4-methylcoumarin-3-acetic acid; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D009834:Oligo-1,6-Glucosidase; D013393:Sucrase